定点突变麻鸡副黏病毒ZH-1株F基因及其鉴定

Identification of the Point Mutation to F Gene of Avian Paramyxovirus Strain ZH-1 Isolated from Ma Chicken

根据麻鸡副黏病毒ZH-1株F基因序列,采用PCR体外定点突变技术,设计2对引物(引物中含有突变位点)。应用重叠PCR延伸法,分3次PCR扩增F基因,从而得到突变后的F基因,命名为F变基因,将其克隆进入pGEM-T载体,并进行PCR、酶切测序鉴定,使突变后的F变基因编码的氨基酸裂解位点为112 Gly-Arg-Gln-Gly-Arg-Leu117,具有弱毒株的特点。

Based on the published F gene sequences of avian paramyxovirus strain ZH-1 isolated from Ma chicken,two pairs of primers containing mutation sites were designed using PCR site-directed mutagenesis in vitro.Appling overlap extension PCR,F gene was PCR amplified for three times to get mutated F genes and then named as F variable genes.PCR products were cloned into pGEM-T Vector,PCR and the positive clone were confirmed by enzyme digestion,the reformed F mutated gene cleavage site amino acid sequence is 112 Gly-Arg-Gln-Gly-Arg-Leu117,which have the character of low virulent strain.